Department of Life Sciences, Graduate School of Arts and Sciences, University of Tokyo, Komaba 3-8-1, Meguro-ku, Tokyo 153-8902, Japan.
DNA Res. 2011 Oct;18(5):393-9. doi: 10.1093/dnares/dsr026. Epub 2011 Jul 29.
Synechocystis sp. PCC 6803 is the most popular cyanobacterial strain, serving as a standard in the research fields of photosynthesis, stress response, metabolism and so on. A glucose-tolerant (GT) derivative of this strain was used for genome sequencing at Kazusa DNA Research Institute in 1996, which established a hallmark in the study of cyanobacteria. However, apparent differences in sequences deviating from the database have been noticed among different strain stocks. For this reason, we analysed the genomic sequence of another GT strain (GT-S) by 454 and partial Sanger sequencing. We found 22 putative single nucleotide polymorphisms (SNPs) in comparison to the published sequence of the Kazusa strain. However, Sanger sequencing of 36 direct PCR products of the Kazusa strains stored in small aliquots resulted in their identity with the GT-S sequence at 21 of the 22 sites, excluding the possibility of their being SNPs. In addition, we were able to combine five split open reading frames present in the database sequence, and to remove the C-terminus of an ORF. Aside from these, two of the Insertion Sequence elements were not present in the GT-S strain. We have thus become able to provide an accurate genomic sequence of Synechocystis sp. PCC 6803 for future studies on this important cyanobacterial strain.
集胞藻 PCC 6803 是最受欢迎的蓝藻菌株，是光合作用、应激反应、代谢等研究领域的标准菌株。1996 年，卡祖萨 DNA 研究所(Kazusa DNA Research Institute)使用该菌株的葡萄糖耐受 (GT) 衍生物进行基因组测序，这为蓝藻研究树立了一个里程碑。然而，不同菌株的数据库序列存在明显差异。因此，我们通过 454 和部分 Sanger 测序分析了另一个 GT 菌株 (GT-S) 的基因组序列。与已发表的卡祖萨株序列相比，我们发现了 22 个可能的单核苷酸多态性 (SNP)。然而，对保存在小等分试样中的卡祖萨株的 36 个直接 PCR 产物进行 Sanger 测序，结果显示在 22 个位点中有 21 个与 GT-S 序列相同，排除了 SNP 的可能性。此外，我们能够将数据库序列中存在的五个分裂开放阅读框组合起来，并去除一个 ORF 的 C 末端。除此之外，GT-S 菌株中不存在两个插入序列元件。因此，我们能够为这个重要的蓝藻菌株的未来研究提供集胞藻 PCC 6803 的准确基因组序列。
